Lung Mechanics: Material Characterization of Pulmonary Constituents for an Experimentally Informed Computational Pipeline.

The lung comprises multiple components including the parenchyma, airways, and visceral pleura, where each constituent displays specific material properties that together govern the whole organ's properties. The structural and mechanical complexity of the lung has historically undermined its comprehensive characterization, especially compared to other biological organs, such as the heart or bones. This knowledge void is particularly remarkable when considering that pulmonary disease is one of the leading causes of morbidity and mortality across the globe. Establishing the mechanical properties of the lung is central to formulating a baseline understanding of its operation, which can facilitate investigations of diseased states and how the lung will potentially respond to clinical interventions. Here, we present established and widely accepted experimental protocols for pulmonary material quantification, specifying how to extract, prepare, and test each type of lung constituent under planar biaxial tensile loading to investigate the mechanical properties, such as physiological stress-strain profiles, anisotropy, and viscoelasticity. These methods are presented across an array of commonly studied species (murine, rat, and porcine). Additionally, we highlight how such material properties may inform the construction of an inverse finite element model, which is central to implementing predictive computational tools for accurate disease diagnostics and optimized medical treatments. These presented methodologies are aimed at supporting research advancements in the field of pulmonary biomechanics and to help inaugurate future novel studies. © 2024 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: General procedures in lung biaxial testing Alternate Protocol 1: Parenchymal-specific preparation and loading procedures Alternate Protocol 2: Airway-specific preparation and loading procedures Alternate Protocol 3: Visceral pleura-specific preparation and loading procedures Basic Protocol 2: Computational analysis.

Visceral pleura mechanics: Characterization of human, pig, and rat lung material properties.

Pulmonary air leaks are amongst the most common complications in lung surgery. Lung sealants are applied to the organ surface and need to synchronously stretch with the visceral pleura, the layer of tissue which encompasses the lung parenchymal tissue. These adhesives are commonly tested on pig and rat lungs, but applied to human lungs. However, the unknown mechanics of human lung visceral pleura undermines the clinical translatability of such animal-tested sealants and the absence of how pig and rat lung visceral pleura compare to human tissues is necessary to address. Here we quantify the biaxial planar tensile mechanics of visceral pleura from healthy transplant-eligible and smoker human lungs for the first time, and further compare the material behaviors to pig and rat lung visceral pleura. Initial and final stiffness moduli, maximum stress, low-to-high strain transition, and stress relaxation are analyzed and compared between and within groups, further considering regional and directional dependencies. Visceral pleura tissue from all species behave isotropically, and pig and human visceral pleura exhibits regional heterogeneity (i.e. upper versus lower lobe differences). We find that pig visceral pleura exhibits similar initial stiffness moduli and regional trends compared to human visceral pleura, suggesting pig tissue may serve as a viable animal model candidate for lung sealant testing. The outcomes and mechanical characterization of these scarce tissues enables future development of biomimetic lung sealants for improved surgical applications. STATEMENT OF SIGNIFICANCE: Surgical lung sealants must synchronously deform with the underlying tissue and with each breath to minimize post-operative air leaks, which remain the most frequent complications of pulmonary intervention. These adhesives are often tested on pig and rat lungs, but applied to humans; however, the material properties of human lung visceral pleura were previously unexplored. Here, for the first time, the mechanics of human visceral pleura tissue are investigated, further contrasting rarely acquired donated lungs from healthy and smoking individuals, and additionally, comparing biaxial planar material characterizations to animal models often employed for pulmonary sealant development. This fundamental material characterization addresses key hindrances in the advancement of biomimetic sealants and evaluates the translatability of animal model experiments for clinical applications.

Pressure-volume mechanics of inflating and deflating intact whole organ porcine lungs.

Pressure-volume curves of the lung are classical measurements of lung function and are impacted by changes in lung structure due to disease or shifts in air-delivery volume or cycling rate. Diseased and preterm infant lungs have been found to show heterogeneous behavior which is highly frequency dependent. This breathing rate dependency has motivated the exploration of multi-frequency oscillatory ventilators to deliver volume oscillation with optimal frequencies for various portions of the lung to provide more uniform air distribution. The design of these advanced ventilators requires the examination of lung function and mechanics, and an improved understanding of the pressure-volume response of the lung. Therefore, to comprehensively analyze whole lung organ mechanics, we investigate six combinations of varying applied volumes and frequencies using ex-vivo porcine specimens and our custom-designed electromechanical breathing apparatus. Lung responses were evaluated through measurements of inflation and deflation slopes, static compliance, peak pressure and volume, as well as hysteresis, energy loss, and pressure relaxation. Generally, we observed that the lungs were stiffer when subjected to faster breathing rates and lower inflation volumes. The lungs exhibited greater inflation volume dependencies compared to frequency dependencies. This study's reported response of the lung to variations of inflation volume and breathing rate can help the optimization of conventional mechanical ventilators and inform the design of advanced ventilators. Although frequency dependency is found to be minimal in normal porcine lungs, this preliminary study lays a foundation for comparison with pathological lungs, which are known to demonstrate marked rate dependency.

Effects of tissue degradation by collagenase and elastase on the biaxial mechanics of porcine airways.

BACKGROUND: Common respiratory illnesses, such as emphysema and chronic obstructive pulmonary disease, are characterized by connective tissue damage and remodeling. Two major fibers govern the mechanics of airway tissue: elastin enables stretch and permits airway recoil, while collagen prevents overextension with stiffer properties. Collagenase and elastase degradation treatments are common avenues for contrasting the role of collagen and elastin in healthy and diseased states; while previous lung studies of collagen and elastin have analyzed parenchymal strips in animal and human specimens, none have focused on the airways to date. METHODS: Specimens were extracted from the proximal and distal airways, namely the trachea, large bronchi, and small bronchi to facilitate evaluations of material heterogeneity, and subjected to biaxial planar loading in the circumferential and axial directions to assess airway anisotropy. Next, samples were subjected to collagenase and elastase enzymatic treatment and tensile tests were repeated. Airway tissue mechanical properties pre- and post-treatment were comprehensively characterized via measures of initial and ultimate moduli, strain transitions, maximum stress, hysteresis, energy loss, and viscoelasticity to gain insights regarding the specialized role of individual connective tissue fibers and network interactions. RESULTS: Enzymatic treatment demonstrated an increase in airway tissue compliance throughout loading and resulted in at least a 50% decrease in maximum stress overall. Strain transition values led to significant anisotropic manifestation post-treatment, where circumferential tissues transitioned at higher strains compared to axial counterparts. Hysteresis values and energy loss decreased after enzymatic treatment, where hysteresis reduced by almost half of the untreated value. Anisotropic ratios exhibited axially led stiffness at low strains which transitioned to circumferentially led stiffness when subjected to higher strains. Viscoelastic stress relaxation was found to be greater in the circumferential direction for bronchial airway regions compared to axial counterparts. CONCLUSION: Targeted fiber treatment resulted in mechanical alterations across the loading range and interactions between elastin and collagen connective tissue networks was observed. Providing novel mechanical characterization of elastase and collagenase treated airways aids our understanding of individual and interconnected fiber roles, ultimately helping to establish a foundation for constructing constitutive models to represent various states and progressions of pulmonary disease.

Biaxial mechanical properties of the bronchial tree: Characterization of elasticity, extensibility, and energetics, including the effect of strain rate and preconditioning.

Distal airways commonly obstruct in lung disease and despite their importance, their mechanical properties are vastly underexplored. The lack of bronchial experiments restricts current airway models to either assume rigid structures, or extrapolate the material properties of the trachea to represent the small airways. Furthermore, past works are exclusively limited to uniaxial testing; investigating the multidirectional tensile loads of both the proximal and distal pulmonary airways is long overdue. Here we present comprehensive mechanical and viscoelastic properties of the porcine airway tree, including the trachea, trachealis muscle, large bronchi, and small bronchi, via measures of elasticity, extensibility, and energetics to explore regional and directional dependencies, cross-examining strain rate and preconditioning effects using planar equibiaxial tensile tests for the first time. We find bronchial regions are notably heterogeneous, where the trachea exhibits greater stiffness, energy loss, and preconditioning sensitivity than the smaller airways. Interestingly, the trachealis muscle is similar to the distal bronchi, despite being anatomically located adjacent to the proximal ring. Tissues are anisotropic and axially stiffer under initial loading, losing more energy with greater stress relaxation circumferentially. Strain rate dependency is also noted, where tissues are more energetically efficient at the faster strain rate, likely attributable to the microstructure. Findings highlight assumptions of homogeneity and isotropy are inadequate, and enable the improvement of aerosol flow and dynamic airway deformation computational predictive models. These results provide much needed fundamental material properties for future explorations contrasting healthy versus diseased pulmonary airway mechanics to better understand the relationship between structure and lung function. STATEMENT OF SIGNIFICANCE: We present comprehensive multiaxial mechanical tensile experiments of the proximal and distal airways via measures of maximum stress, initial and ultimate moduli, strain and stress transitions, hysteresis, energy loss, and stress relaxation, and further assess preconditioning and strain rate dependencies to examine the relationship between lung function and structure. The mechanical response of the bronchial tree demonstrates significant anisotropy and heterogeneity, even within the tracheal ring, and emphasizes that contrary to past studies, the behavior of the proximal airways cannot be extended to distal bronchial tree analyses. Establishing these material properties is critical to advancing our understanding of airway function and in developing accurate computational simulations to help diagnose and monitor pulmonary diseases.

Positive- and Negative-Pressure Ventilation Characterized by Local and Global Pulmonary Mechanics.

Rationale: There is continued debate regarding the equivalency of positive-pressure ventilation (PPV) and negative-pressure ventilation (NPV). Resolving this question is important because of the different practical ramifications of the two paradigms. Objectives: We sought to investigate the parallel between PPV and NPV and determine whether or not these two paradigms cause identical ventilation profiles by analyzing the local strain mechanics when the global tidal volume (Vt) and inflation pressure was matched. Methods: A custom-designed electromechanical apparatus was used to impose equal global loads and displacements on the same ex vivo healthy porcine lung using PPV and NPV. High-speed high-resolution cameras recorded local lung surface deformations and strains in real time, and differences between PPV and NPV global energetics, viscoelasticity, as well as local tissue distortion were assessed. Measurements and Main Results: During initial inflation, NPV exhibited significantly more bulk pressure-volume compliance than PPV, suggestive of earlier lung recruitment. NPV settings also showed reduced relaxation, hysteresis, and energy loss compared with PPV. Local strain trends were also decreased in NPV, with reduced tissue distortion trends compared with PPV, as revealed through analysis of tissue anisotropy. Conclusions: Apparently, contradictory previous studies are not mutually exclusive. Equivalent changes in transpulmonary pressures in PPV and NPV lead to the same changes in lung volume and pressures, yet local tissue strains differ between PPV and NPV. Although limited to healthy specimens and ex vivo experiments in the absence of a chest cavity, these results may explain previous reports of better oxygenation and less lung injury in NPV.

Associating local strains to global pressure-volume mouse lung mechanics using digital image correlation.

Pulmonary diseases alter lung mechanical properties, can cause loss of function, and necessitate use of mechanical ventilation, which can be detrimental. Investigations of lung tissue (local) scale mechanical properties are sparse compared to that of the whole organ (global) level, despite connections between regional strain injury and ventilation. We examine ex vivo mouse lung mechanics by investigating strain values, local compliance, tissue surface heterogeneity, and strain evolutionary behavior for various inflation rates and volumes. A custom electromechanical, pressure-volume ventilator is coupled with digital image correlation to measure regional lung strains and associate local to global mechanics by analyzing novel pressure-strain evolutionary measures. Mean strains at 5 breaths per minute (BPM) for applied volumes of 0.3, 0.5, and 0.7 ml are 5.0, 7.8, and 11.3%, respectively, and 4.7, 8.8, and 12.2% for 20 BPM. Similarly, maximum strains among all rate and volume combinations range 10.7%-22.4%. Strain values (mean, range, mode, and maximum) at peak inflation often exhibit significant volume dependencies. Additionally, select evolutionary behavior (e.g., local lung compliance quantification) and tissue heterogeneity show significant volume dependence. Rate dependencies are generally found to be insignificant; however, strain values and surface lobe heterogeneity tend to increase with increasing rates. By quantifying strain evolutionary behavior in relation to pressure-volume measures, we associate time-continuous local to global mouse lung mechanics for the first time and further examine the role of volume and rate dependency. The interplay of multiscale deformations evaluated in this work can offer insights for clinical applications, such as ventilator-induced lung injury.

Developing a Lung Model in the Age of COVID-19: A Digital Image Correlation and Inverse Finite Element Analysis Framework.

Pulmonary diseases, driven by pollution, industrial farming, vaping, and the infamous COVID-19 pandemic, lead morbidity and mortality rates worldwide. Computational biomechanical models can enhance predictive capabilities to understand fundamental lung physiology; however, such investigations are hindered by the lung's complex and hierarchical structure, and the lack of mechanical experiments linking the load-bearing organ-level response to local behaviors. In this study we address these impedances by introducing a novel reduced-order surface model of the lung, combining the response of the intricate bronchial network, parenchymal tissue, and visceral pleura. The inverse finite element analysis (IFEA) framework is developed using 3-D digital image correlation (DIC) from experimentally measured non-contact strains and displacements from an ex-vivo porcine lung specimen for the first time. A custom-designed inflation device is employed to uniquely correlate the multiscale classical pressure-volume bulk breathing measures to local-level deformation topologies and principal expansion directions. Optimal material parameters are found by minimizing the error between experimental and simulation-based lung surface displacement values, using both classes of gradient-based and gradient-free optimization algorithms and by developing an adjoint formulation for efficiency. The heterogeneous and anisotropic characteristics of pulmonary breathing are represented using various hyperelastic continuum formulations to divulge compound material parameters and evaluate the best performing model. While accounting for tissue anisotropy with fibers assumed along medial-lateral direction did not benefit model calibration, allowing for regional material heterogeneity enabled accurate reconstruction of lung deformations when compared to the homogeneous model. The proof-of-concept framework established here can be readily applied to investigate the impact of assorted organ-level ventilation strategies on local pulmonary force and strain distributions, and to further explore how diseased states may alter the load-bearing material behavior of the lung. In the age of a respiratory pandemic, advancing our understanding of lung biomechanics is more pressing than ever before.

Novel Mechanical Strain Characterization of Ventilated ex vivo Porcine and Murine Lung using Digital Image Correlation.

Respiratory illnesses, such as bronchitis, emphysema, asthma, and COVID-19, substantially remodel lung tissue, deteriorate function, and culminate in a compromised breathing ability. Yet, the structural mechanics of the lung is significantly understudied. Classical pressure-volume air or saline inflation studies of the lung have attempted to characterize the organ's elasticity and compliance, measuring deviatory responses in diseased states; however, these investigations are exclusively limited to the bulk composite or global response of the entire lung and disregard local expansion and stretch phenomena within the lung lobes, overlooking potentially valuable physiological insights, as particularly related to mechanical ventilation. Here, we present a method to collect the first non-contact, full-field deformation measures of ex vivo porcine and murine lungs and interface with a pressure-volume ventilation system to investigate lung behavior in real time. We share preliminary observations of heterogeneous and anisotropic strain distributions of the parenchymal surface, associative pressure-volume-strain loading dependencies during continuous loading, and consider the influence of inflation rate and maximum volume. This study serves as a crucial basis for future works to comprehensively characterize the regional response of the lung across various species, link local strains to global lung mechanics, examine the effect of breathing frequencies and volumes, investigate deformation gradients and evolutionary behaviors during breathing, and contrast healthy and pathological states. Measurements collected in this framework ultimately aim to inform predictive computational models and enable the effective development of ventilators and early diagnostic strategies.

Introducing a Custom-Designed Volume-Pressure Machine for Novel Measurements of Whole Lung Organ Viscoelasticity and Direct Comparisons Between Positive- and Negative-Pressure Ventilation.

Asthma, emphysema, COVID-19 and other lung-impacting diseases cause the remodeling of tissue structural properties and can lead to changes in conducting pulmonary volume, viscoelasticity, and air flow distribution. Whole organ experimental inflation tests are commonly used to understand the impact of these modifications on lung mechanics. Here we introduce a novel, automated, custom-designed device for measuring the volume and pressure response of lungs, surpassing the capabilities of traditional machines and built to range size-scales to accommodate both murine and porcine tests. The software-controlled system is capable of constructing standardized continuous volume-pressure curves, while accounting for air compressibility, yielding consistent and reproducible measures while eliminating the need for pulmonary degassing. This device uses volume-control to enable viscoelastic whole lung macromechanical insights from rate dependencies and pressure-time curves. Moreover, the conceptual design of this device facilitates studies relating the phenomenon of diaphragm breathing and artificial ventilation induced by pushing air inside the lungs. System capabilities are demonstrated and validated via a comparative study between ex vivo murine lungs and elastic balloons, using various testing protocols. Volume-pressure curve comparisons with previous pressure-controlled systems yield good agreement, confirming accuracy. This work expands the capabilities of current lung experiments, improving scientific investigations of healthy and diseased pulmonary biomechanics. Ultimately, the methodologies demonstrated in the manufacturing of this system enable future studies centered on investigating viscoelasticity as a potential biomarker and improvements to patient ventilators based on direct assessment and comparisons of positive- and negative-pressure mechanics.

Synthesis, characterization, and cytocompatibility of yttria stabilized zirconia nanopowders for creating a window to the brain.

Transparent cranial window to the brain is highly desirable for brain therapies because such cranial implant would allow for continuous monitoring of brain disorders and long-term delivery of photodynamic therapy into the brain without repeated surgeries for opening skull. Nanostructured yttria-stabilized zirconia (YSZ) is a potential candidate for the window to the brain application because of its promising mechanical and optical properties. In this study, a new process using aerosol spray pyrolysis was established for synthesizing 6-7 nm YSZ nanopowders with precisely controlled compositions. YSZ nanopowders with 3 M ratios of yttria to zirconia, specifically 3, 6, and 8% yttria in zirconia (referred to as 3YSZ, 6YSZ, and 8YSZ, respectively) were synthesized and characterized. The size, structure, and composition of the produced YSZ nanoparticles are highly controllable and scalable. The in vitro cytocompatibility of the YSZ nanoparticles with bone marrow mesenchymal stem cells (BMSCs) was investigated using a direct exposure culture method for cranial implant applications. Nondoped ZrO2 and commercially available 8YSZ (named as C_8YSZ) served as controls for the in vitro cell studies. BMSCs exhibited normal morphology when cultured with the YSZs of 3 M ratios in the concentrations of 10 mM, 30 mM, and 60 mM, as well as ZrO2 and C_8YSZ controls. The BMSCs cultured with 3YSZ and 6YSZ showed no statistical differences in cell adhesion density when compared with the ZrO2 and C_8YSZ controls at respective concentrations of 10-60 mM. The possible release of YSZ nanoparticles from cranial window implants should be carefully considered and further studied.